The ATRX Knockout HT29 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal population of cells featuring disruption of the ATRX gene in the HT29 colorectal adenocarcinoma background. This product provides a heterogeneous knockout cell pool generated through targeted CRISPR/Cas9-mediated gene disruption, offering a versatile model for investigating ATRX-dependent cellular processes without the constraints of single-cell clonal selection. The polyclonal nature captures a spectrum of editing events across the population, facilitating functional studies that mimic the heterogeneity observed in tumor microenvironments in vivo.
HT29 is a well-characterized human colorectal adenocarcinoma cell line originally derived from a primary tumor of a 44-year-old female. These cells form polarized epithelial monolayers with intact tight junctions and retain key features of colonic epithelium, making them extensively used for studies of intestinal barrier function, colorectal cancer biology, and drug transport. The HT29 background provides a physiologically relevant epithelial context for the ATRX knockout, enabling mechanistic investigation within the framework of an adenocarcinoma model that reflects both differentiation capacity and malignant characteristics.
ATRX encodes a chromatin remodeler that acts as a histone H3.3 chaperone in complex with DAXX, mediating deposition of histone H3.3 at telomeres, pericentric heterochromatin, and other regions. ATRX functions downstream of DNA damage response signaling and cell cycle cues, and interacts with HP1?? and PML nuclear bodies to promote heterochromatin assembly. Disruption of ATRX impairs H3.3 incorporation at telomeres, triggering telomere dysfunction, activation of the ALT pathway, and genomic instability. ATRX loss also dysregulates transcriptional programs through its chromatin remodeling role. Key pathway components include DAXX, H3F3A (encoding H3.3), TERRA non-coding RNA, and telomerase subunits (TERT, TERC).
In the context of HT29 colorectal adenocarcinoma cells, ATRX knockout provides a powerful model to dissect the molecular consequences of ATRX deficiency relevant to colorectal cancer progression and other malignancies such as glioma and neuroendocrine tumors. ATRX mutations are recurrently observed in cancers that employ ALT, and the HT29 background with its adenocarcinoma phenotype allows the study of how ATRX loss cooperates with existing oncogenic lesions. The knockout model enables exploration of telomere maintenance mechanisms, chromatin remodeling defects, and DNA damage response pathways in an epithelial cancer setting, providing insights into tumor biology and potential therapeutic vulnerabilities.
This ATRX knockout polyclonal cell product is suited for diverse research applications. Western blot analysis confirms ATRX protein loss, and telomere FISH detects ALT-associated telomeric foci. Immunofluorescence co-staining for histone H3.3 and PML bodies assesses disrupted chromatin assembly, and ??H2AX staining reveals DNA double-strand break accumulation. With cell proliferation assays and RT-qPCR for gene expression, these cells support studies of ALT mechanisms, chromatin remodeling in cancer, drug sensitivity screening, colorectal cancer biology, and ATR-X syndrome modeling. For additional technical specifications, contact Ascent Research.
